APN1的原核表达及其与Cry1Ac 蛋白体外结合

摘要

Binding assay between Cry1Ac toxin and BBMVs from Helicoverpa armigera was analyzed in this study. The full-length gene and the fragments of apn1 were cloned and expressed in Escherichia coli Rosetta (DE3) strain.The binding between full-length/fragments of APN1 expressed in E .coli and Cry1Ac toxin was performed by Ligand blotting.The results showed that BBMVs from H .armigera could bind Cry1Ac toxin;the full-length APN1 and APN1 fragments H1 (33-257),H2(258-547),and H3(548-798)expressed in prokaryotic system could bind Cry1Ac toxin.It was the first report that H3 (548-798)could bind with Cry1Ac.The results may provide a new clue for functional studies on other Bt toxin receptors in vitro .%本研究分析了 Cry1Ac 蛋白与棉铃虫 BBMVs 的结合情况；并将棉铃虫 apn1基因的全长和片段在大肠杆菌Rosetta（DE3）中进行表达，利用 Ligand blotting 技术分析原核表达的 APN1全长及4个片段与 Cry1Ac 蛋白的结合情况。结果表明：Cry1Ac 蛋白可以与棉铃虫的 BBMVs 结合；在大肠杆菌中表达的棉铃虫受体蛋白 APN1的全长、片段 H1（33～257）、H2（258～547）和 H3（548～798）可以与 Cry1Ac 蛋白结合，其中片段 H3是首次发现可以与Cry1Ac 蛋白结合。本研究的结果也为其他 Bt 蛋白受体的体外功能研究提供新的借鉴。